LIPID ANALYSIS OF FILLETS FROM GIANT GRENADIER

(ALBATROSSIA PECTORALIS), ARROW-TOOTH FLOUNDER

(ATHERESTHES STOMIAS), PACIFIC COD (GADUS
MACROCEPHALUS) AND WALLEYE POLLOCK
(THERAGRA CHALCOGRAMMA)

A.C.M. OLIVEIRA1,3 and P.J. BECHTEL2

1
Fishery Industrial Technology Center
University of Alaska Fairbanks
118 Trident Way, Kodiak, AK 99615
2
USDA–ARS, Seafood Laboratory
233 O’Neill Building, University of Alaska
Fairbanks, AK 99775

Accepted for Publication March 29, 2005

ABSTRACT

In Alaska, walleye pollock (WP [Theragra chalcogramma]) and Pacific

cod (PC [Gadus macrocephalus]) are harvested in large amounts. There are
also large biomasses of arrow-tooth flounder (ATF [Atheresthes stomias]) and
giant grenadier (GG [Albatrossia pectoralis]) available in Alaskan waters;
however, these fish are underutilized. The objective of this study was to deter-
mine lipid content, lipid class distribution and fatty acid (FA) profiles of fillets
from these four fish species. Fat content and percent triacylglycerides (TAG) in
ATF was significantly higher than in other species, and percent phospholipids
(PL) was lower (P ⬍ 0.05). PC had the highest levels of omega-3 (w-3) FA,
whereas ATF had the lowest. The ratio of w-3/omega-6 (w-6) FA was signifi-
cantly higher in ATF than in other species. Percent polyunsaturated and
saturated FA’s were equivalent in ATF; however, other species showed at least
twice as much polyunsaturated FA as saturated FA. This study provided com-
parisons of the lipid characteristics in the two most abundant white fish
harvested in Alaska and two other abundant but underutilized species.

INTRODUCTION

Over 60% of the fish harvested from U.S. waters is used for human
consumption and comes from Alaskan waters. In Alaska, the fish harvested in
3
Corresponding author. TEL: (907) 486-1530; FAX: (907) 486-1540; EMAIL: ffamo@uaf.edu

Journal of Muscle Foods 17 (2006) 20–33. All Rights Reserved.

20 © 2006, The Author(s)
Journal compilation © 2006, Blackwell Publishing
 LIPID ANALYSIS OF FISH FILLETS 21

the largest amounts are walleye pollock (WP [Theragra chalcogramma]) and
Pacific cod (PC [Gadus macrocephalus]). WP commercial catches exceeded
1,000,000 t in 2000 while PC catches in the same year were approximately
225,000 t (Crapo and Bechtel 2003). There are large biomasses of arrow-tooth
flounder (ATF [Atheresthes stomias]) and giant grenadier (GG [Albatrossia
pectoralis]) that are underutilized. The latter, also called rattail, is a nontar-
geted catch in the deep water trawl fishery in Alaska (Crapo et al. 1999). Low
(2003) recently reported that the long-term annual potential yield of ATF in the
Bering Sea–Aleutians region is 137,000 t.
During the 1990s, there was remarkable growth in the abundance of ATF
and GG in Alaskan waters (Alaska Fishery Science Center [AFSC] 2001).
However, these two species are not commercially harvested because of intrin-
sic quality issues regarding the edible muscle. Babbit et al. (1993) reported the
presence of a protease in ATF which causes the flesh to become soft during
cooking, rendering it unmarketable. Crapo et al. (1999) attributed low meat
yields and flesh softness of GG to its unusually high moisture content (91.0%)
combined with a low protein content (6.8%).
There is much interest in the beneficial effects on human health that is
related to consumption of marine lipids. In addition, there is a growing need to
find commercial uses for underutilized but abundant marine cold-water fish
species. Therefore, the objective of this study was to determine and compare
the lipid content, lipid classes (triacylglycerides, diacylglycerides, monoacylg-
lycerides, phospholipids, sterols and free fatty acids; TAG, DAG, MAG, PL,
ST and FFAs, respectively) and the FA composition of fillets from WP, PC,
ATF and GG.

MATERIALS AND METHODS

Fish Sampling
Triplicate samples of WP and PC dressed fillets were obtained during a
1-week period in February 2001 from Alaskan fish processors in Kodiak,
Alaska, that utilized automated heading, filleting and fillet-skinning opera-
tions. Samples for each species were taken during three separate days from a
minimum of two different processing plants in order to allow sampling of fish
harvested by different fishing vessels. Dressed fillets were prepared from fresh
harvested GG and ATF that became available as bycatch during April 2002 in
the University of Alaska Fishery Industrial Technology Center’s pilot plant
located in Kodiak, Alaska. The fish were collected from the plants and filleted
immediately or frozen in a -30C blast freezer overnight for processing on the
following day. There were three replicates for each species, totaling 12
samples. All samples were ground and stored at -70C until solvent extraction.
22 A.C.M. OLIVEIRA and P.J. BECHTEL

Lipid Extraction
The lipid extraction procedure was adapted from Radin (1981). Accord-
ingly, 18 mL of a 2:3 solution of isopropyl alcohol (IPA)/hexane (99.9%
purity; Burdick & Jackson, Muskegon, MI) was added per 1 g of sample. The
mixture was blended to a slurry for 1 min under nitrogen (N2) gas. The
contents were filtered under vacuum through a 150-mL Pyrex sintered funnel
lined using a Whatman #2 filter paper 70 mm in diameter (both from VMR
International, Brisbane, CA). The filtrate was collected into a 500-mL side-
arm Erlenmeyer flask, and two additional portions (3 and 2 mL/g sample) of
the IPA/hexane (2:3) solvent mixture were used to extract traces of tissue lipids
retained on the filter paper. The filtrate was decanted into a 500-mL pear-
shaped flask and the solvent removed at 49C on a rotary evaporator (Büchi
Rotavapor R-205, Westbury, NY) to a lipid–solvent volume of approximately
5 mL. The oil and remaining solvent were decanted into a 50-mL Nalgene
screw-top centrifuge tube (VWR International) and two 10-mL portions of
hexane were used to rinse the same flask and added to the centrifuge tube.
Ten milliliters of distilled water was added to the centrifuge tube and mixed
(vortex) for 1–2 min. The mixture was centrifuged for 20 min at 3400 rpm
using an IEC Centra CL2 centrifuge (IEC, Needham Heights, MA). The top
layer was transferred into a small Erlenmeyer flask containing 2.5–3.0 g of
anhydrous sodium sulfate of 99+% purity (Aldrich, Milwaukee, WI) for about
30 min and then filtered through a Whatman #1 filter paper (VWR Interna-
tional) to remove water before transferring to a 50-mL pear-shaped flask.
Another 10 mL of hexane was added to the remaining contents of the centri-
fuge tube and repeated the procedure. Solvent was removed using a rotatory
evaporator and the lipids were transferred into a preweighed 10-mL screw-top
amber vial. The remaining solvent was removed under a N2 gas stream. The
weights were recorded and percent lipids determined gravimetrically. Oils
were flushed with N2 and stored in amber vials at -80C until analyses.

Preparation of Fatty Acid Methyl Esters

Fatty acid methyl esters (FAME) were prepared using the method of
Maxwell and Marmer (1983). Approximately 20 mg of oil was placed into a
10-mL round-bottom screw-cap centrifuge tube and the exact weight recorded.
The lipids were dissolved at ambient temperature in 1.9 mL of isooctane (99%
grade, Sigma, St. Louis, MO) and 100 mL of a solution of 10-mg/mL tri-
cosanoic acid methyl ester (C23:0) (Supelco, Bellefonte, PA) in isooctane and
then 200 mL of 2-N KOH in methanol (1.12 g/10 mL) was added to the
centrifuge tube. Contents were mixed for 60 s, centrifuged for 3 min at
3400 rpm and the lower layer discarded. This procedure was repeated twice
using 0.5 mL of a saturated solution of ammonium acetate in water, followed
 LIPID ANALYSIS OF FISH FILLETS 23

by 0.5 mL of deionized water. Methyl esters in isooctane were then dried by

adding 200 to 300 mg of anhydrous sodium sulfate for 20 min which was
removed by centrifugation at 3400 rpm for 20 min. The FAMEs were trans-
ferred into 1.5-mL snap-cap gas chromatography (GC) amber vials (Agilent
Technologies, Wilmington, DE) for chromatographic analysis.

GC Analysis
An Agilent Technologies GC model 6850 fitted with a DB225
(30 m ¥ 0.25-mm i.d. with 0.25-mm film) capillary column (Agilent Technolo-
gies) was used for FA analysis. Data were collected and analyzed using the GC
ChemStation program (Rev.A.08.03 [847] 1990–2000; Agilent Technologies).
Helium was used as carrier gas at constant flow of 1.0 mL/min. The injector
and detector temperatures were 250C. The split ratio was 25:1 and the oven
programming was 140 to 220C at a rate of 3C/min for a total run time of
47 min. An autosampler performed the GC injections of standards and
samples. The injection volume was 1 ml. The ChemStation enhanced integrator
program was used to integrate the chromatogram peaks. All standards used in
the identification of peaks were purchased from Supelco and included Supelco
37, Bacterial Acid Methyl Esters Mix and Marine Oils #1 and #3. The samples
were run in duplicate. Cod-liver oil was used as a secondary reference standard
(Ackman and Burgher 1965).

Lipid Classes Analysis

An Iatroscan TLC/FID Analyzer model MK-6s (Iatron Laboratories Inc.,
Tokyo, Japan) was used to determine percent areas for TAG, DAG, MAG, free
FAs, ST and PL in the oil samples. The materials and methods used for
lipid-class analyses were adapted from Whitsett et al. (1986), Parrish (1987)
and Ackman et al. (1990). The hydrogen flow rate was 60 mL/min, the airflow
was 1.6 L/min and the scan time was 30 s. Six standards (Sigma) were used to
identify the lipid classes found in the fish oil and included cholesterol (ST),
triolein (TG), oleic acid (FFA), phosphatidylcholine (PL), diolein (DG) and
monopalmitoylglycerol (MG). Cod-liver oil (Marine Bio Products, Quincy,
MA) and commercially available fish oils were used during the test phase to
determine the best sample concentration to be spotted on the rods. Peaks were
integrated using the software Peak Simple Program (version 2.83, 6 channels,
SRI Instruments, Torrance, CA). The solvent system used was a mixture of
hexane : ethyl ether : formic acid at the ratio of 80.0:25.0:1.2 (Parrish 1987).
The solvent tank (Shell-U.S.A. Inc., Fredericksburg, VA) was lined with filter
paper and 60 mL of solvent. The equilibration time of the closed system was
20 min. Chromarods-SIII (Iatron Laboratories, Inc.) were cleaned for sample
24 A.C.M. OLIVEIRA and P.J. BECHTEL

spotting by performing consecutive origin scans at a speed of 50 s until a flat

baseline was recorded. A final blank scanning at a speed of 30 s finished
conditioning the rods for spotting. A 1-mL aliquot of a 10.0-mg/mL solution of
oil in chloroform was spotted in each rod. For each set of 10 rods, 2 oil samples
were spotted in quadruplicate and a 1-mL aliquot of a composite standard
mixture containing about 2.5 mg/mL of each standard in chloroform was
spotted on duplicate rods. The rods were suspended in the solvent chamber for
10 min then lowered into the solvent front for 30 min (Ackman et al. 1990).
These were oven-dried for 3 min at 110C and scanned promptly (Whitsett
et al. 1986). When necessary, rods were cleaned overnight by soaking in a
concentrated mixture of chromic and sulfuric acid. Then these were rinsed
with distilled water and soaked for 3 h in 5% ammonium hydroxide. The rods
were then rinsed several times with distilled water followed by a final acetone
rinse and oven-dried for 1 to 2 h. Clean rods were scanned once and stored in
a 30% humidity chamber (Parrish 1987).

Statistical Analysis
The weighted means are derived from an analysis of variance run on
Statistica (version 6.0, StatSoft Inc., Tulsa, OK). For tests of statistical sig-
nificance between species, the data were subjected to Duncan’s posthoc test
with significance set at P ⬍ 0.05.

RESULTS AND DISCUSSION

Lipid Content
Table 1 lists the lipid content for the four fish species studied. The highest
lipid content was in ATF at 3.5% (P ⬍ 0.05). This value is in agreement with
the 4.52–0.78% range for lipid content reported for this species (Stansby
1976). The percent lipids determined for WP (0.6%) was similar to the low end
of oil content reported by Stansby (1976) and it is also in agreement with the
average values reported for Bering Sea WP of 0.7% (Krzynowek and Murphy
1987) and 0.8% (U.S. Department of Agriculture–Agricultural Research
Service 2004a). The average value reported for lipid content in WP collected
in Alaska during the summers of 1993 and 1994 was 1.44% (Van Pelt et al.
1997). The lipid content of PC fillets was similar to WP (P ⬍ 0.05) and agrees
with the 0.66 and 0.63% oil content reported by Krzynowek and Murphy
(1987) and the U.S. Department of Agriculture–Agricultural Research Service
(2004b), respectively. The oil content of PC caught during the summer months
was 0.98% as reported by Van Pelt et al. (1997). The lipid content of GG was
 LIPID ANALYSIS OF FISH FILLETS 25

TABLE 1.
LIPID CLASS ANALYSES OF EDIBLE MUSCLE (PERCENT AREA)

Species Lipids TAG FFA DAG ST MAG PL

(SD) (SD) (SD) (SD) (SD) (SD) (SD)

Giant grenadier 0.21A 7.71A 12.53A 5.08A 0.48A 0.48A 73.72A

(0.02) (2.10) (2.70) (0.43) (0.24) (0.21) (1.69)
Arrow-tooth 3.51B 80.22B 0.43B 1.10B 0.46A 0.45A 15.82B
flounder (0.62) (2.98) (0.11) (0.51) (0.16) (0.18) (1.32)
Pacific cod 0.59A 2.87C 16.92A 5.89AC 0.45A 0.68A 73.18A
(0.01) (0.12) (2.72) (0.62) (0.14) (0.17) (3.47)
Walleye pollock 0.60A 3.03C 20.65A 7.38C 0.72A 0.77A 67.45A
(0.08) (1.47) (7.66) (1.36) (0.12) (0.10) (7.99)

Different superscript letters show significant differences by column at P ⬍ 0.05.

TAG, triacylglycerides; FFA, free fatty acids; DAG, diacylglycerides; ST, sterols; MAG, monoacylg-
lycerides; PL, phospholipids; SD, standard deviation of the mean.

the lowest of all species at 0.2% (P ⬍ 0.05) which agrees with the value
reported for the oil content of fish harvested in prime condition (Crapo et al.
1999). Crapo et al. (1999) reported that, at late stages of spawning, GG fillets
show a 50% decrease in lipid content. The lipid content of other members of
the roughhead grenadiers was reported at 0.47%, while percent lipids for
roundnose grenadier were estimated at 0.69% (Botta and Shaw 1976).

Lipid Classes Distribution

Results for lipid classes are reported in percent area with sum of areas for
all peaks of interest equivalent to 100%. Calculations of lipid classes as
percent total lipids (w/w basis) were not included because of difficulties in
obtaining reliable standard curves for very low concentrations of specific lipid
classes, which corresponded to small areas. Pure compounds, listed above,
were spotted in quadruplicate at four increasing concentrations from 1 to
10 mg/mL with a 1-mL load. The slopes determined for the standards TG, FFA,
DG and MG were very similar and corresponded to 5.35, 4.24, 5.04 and 4.81,
respectively. However, the slopes determined for ST and PL were much higher,
at 7.2 and 13.7, respectively. According to Parrish (1987), ST is a high
response compound presenting a higher slope than TAG, DAG, MAG, FFA
and PL, which were defined as “low” response compounds. In our study
however, PL had the highest slope of all lipid classes investigated. It should be
noted that even though we have reported the differences in FID response
among some of the lipid classes, the mentioned difficulties led us to present the
results from lipid classes in percent areas as integrated by the instrument.
As shown in Table 1, ST and MAG content were both below 1%, and not
significantly different among the four species. The percent ST determined for
26 A.C.M. OLIVEIRA and P.J. BECHTEL

WP agrees with the reported value for ST content in pollock-edible flesh at

75% (Krzynowek and Murphy 1987). The system used 1,3 DAG coelutes with
the ST peak separately from the 1,2 DAG peak, which is designated as DAG
in Table 1. The DAG content (1,2 DAG) was variable among species, ranging
from 1.0 to 7.4%, with ATF showing the lowest abundance and the remaining
species varying from 5.1 to 7.4%. The PL represented about 70% of the area
for all species, except ATF. ATF fillets, which had the highest fat content, were
highest in percent triglycerides and lowest in PL. Whitsett et al. (1986)
reported that PL are the major lipid components of lean tissue while TAG are
predominant in tissues with higher fat content. The FFA content was higher for
WP, PC and GG than AFT, although it should be noted these fillet values were
much lower than FFA values from stored tissues with more lipase activity, such
as the viscera (Oliveira and Bechtel 2005). The WP and PC samples were
collected and stored for approximately 10 months at -70C before extraction
and analysis. The ATF and GG fillet samples were stored for less than 2 weeks
prior to extraction and analysis. The same laboratory procedures and instru-
ments were used for all samples and standards to insure reliability in compar-
ing results. The FFA values for PC and WP could be partially because of lipase
activity during sample storage; however, GG had a percent FFA similar to both
WP and PC. Another explanation for this finding is related to the fat content of
the tissue. We have observed in a previous study (Oliveira and Bechtel 2005)
that percent FFA tends to be higher in tissue with lower lipid content, espe-
cially tissues having lipid levels below 1%. Percent FFA determined by Iatros-
can on Chromarods-SIII was reported for Spanish sardines (SS), chub
mackerel (CM) and thread herring (Hale and Brown 1983). Lipid content in
these three fish species varied from 3.2% (SS) to 2.7% (CM), and FFA content
from 4.0% (SS) to 7.0% (CM).

FAs Profile
Table 2 presents the overall results of FAME analysis as percent of total
FA in the sample. There were significant differences in the percent saturated
fatty acids (SFA); however, only a 4% difference between lowest and highest
concentration of SFA was found. The percent SFA in cod of unidentified origin
reported by Pearson (1977) was 32%; while Kovacs and Ackman (1978)
reported values of 0.1 g/100 g of tissue. Percent monounsaturated fatty acids
(MUSA) were highly variable among species, with ATF having over twice the
percent (39.9%) determined for PC (P ⬎ 0.05), and values for both GG and
WP were similar but significantly different from both ATF and PC values. On
the other hand, percent polyunsaturated fatty acids (PUFAs) were lowest in
ATF at about half the amount determined for other species (P ⬎ 0.05). GG, PC
and WP had over 40% of their total fatty acids as PUFA. These findings were
 LIPID ANALYSIS OF FISH FILLETS 27

TABLE 2.
FATTY ACID METHYL ESTER ANALYSIS SUMMARY (PERCENT TOTAL FATTY ACIDS)

Species SFA MUSA PUFA P/S w-3 w-6 w-3/w-6

(SD) (SD) (SD) (SD) (SD) (SD) (SD)

Giant grenadier 27.27AB 27.40A 43.31A 1.59A 44.14A 3.76A 11.74A

(0.84) (1.85) (1.42) (0.02) (3.32) (0.39) (0.52)
Arrow-tooth 25.72B 38.89B 26.02B 1.01B 25.35B 2.15B 11.79A
flounder (1.34) (3.29) (2.00) (0.06) (1.83) (0.16) (0.45)
Pacific cod 23.40C 17.85C 54.80C 2.34C 50.26C 3.88A 12.95A
(0.43) (2.49) (2.30) (0.03) (2.24) (0.28) (1.15)
Walleye pollock 27.55A 25.12A 46.06A 1.67A 43.57A 2.49B 17.50B
(0.23) (1.99) (2.23) (0.03) (2.23) (0.23) (1.85)

Different superscript letters show significant differences by column at P ⬍ 0.05.

SFA, saturated fatty acids; MUSA, monounsaturated fatty acids; PUFA, polyunsaturated fatty acids;
P/S, % PUFA/% SFA; w-3, total omega-3 fatty acids; w-6, total omega-6 fatty acids; w-3/ w-6, % total
omega-3 fatty acids/total % omega-6 fatty acids; SD, standard deviation of the mean.

reflected in the % PUFA/% SFA (P/S) ratios, with ATF fillets having a signifi-
cantly lower P/S ratio and PC having the highest ratio. Omega-3 (w-3) content
as percent of total fatty acids followed the same pattern as PUFA, with ATF
having the lowest percent w-3 (25.4%) and PC the highest (50.3%). Percent
PUFA and w-3 for PC agree with values previously reported by Stansby
(1976). The ratio of w-3/omega-6 (w-6) was highest (P ⬎ 0.05) for WP fillets
(17.5); the ratios for all other species did not differ, ranging from 11.7 to 13.0.
The percent SFA, MUSA and PUFA of GG and WP fillets were similar.
Both species had very low lipid contents. PC fillets also had a very low lipid
content of 0.6%; however, the percent PUFA was higher because of increased
concentrations of docosahexaenoic acid (DHA) and decreased concentrations
of MUSA such as vaccenic acid (C18:1w7). ATF fillets had a lipid content over
five times greater than GG, WP and PC; over 80% of the ATF lipid was TAGs.
The large MUSA values found in ATF were because of increased concentra-
tions of C16:1w7, FFA cis isomer (C18:1w9) and other MUSAs. The large
reduction in ATF–PUFA values when compared to PUFA values from the other
species was attributed in large part to the reduction in w-3 FAs, especially
DHA. It is of interest to note that although ATF has a reduced concentration of
DHA, the levels of MUSAs are very high for a marine fish oil.
Table 3 takes into account total fat content of fillets and shows mg of FA
class per 100 g of edible muscle. Lipid content of the fillets varied from 0.2%
in GG to 3.5% in ATF, and as expected, mg of saturated fat was lowest for GG
and highest for ATF. Similar patterns were seen for PUFA content, with ATF
having the greatest amount, followed by PC, WP and GG. The PUFA content
for PC (0.33 g/100 g of tissue) was slightly higher than the 0.20 g/100 g of
28 A.C.M. OLIVEIRA and P.J. BECHTEL

TABLE 3.
FATTY ACID METHYL ESTER ANALYSIS SUMMARY (mg/100 g of tissue)

Species SFA MUSA PUFA w-3 w-6 EPA DHA

(SD) (SD) (SD) (SD) (SD) (SD) (SD)

Giant grenadier 58.20A 55.63A 96.61A 94.27A 8.04A 27.70A 49.75A

(6.34) (5.79) (9.91) (12.63) (1.39) (2.55) (2.80)
Arrow-tooth 899.85B 1275.71B 1053.96B 882.94B 74.87B 439.47B 264.20B
flounder (127.74) (31.57) (211.93) (87.18) (8.67) (27.98) (39.98)
Pacific cod 138.06A 101.28C 328.53C 296.64C 22.91C 79.04C 207.59B
(1.61) (17.77) (21.92) (16.72) (1.93) (6.40) (7.46)
Walleye pollock 166.15A 142.88D 286.06AC 263.40C 15.13AC 85.00C 175.29C
(21.64) (13.92) (40.02) (41.78) (3.25) (8.47) (12.94)

Different superscript letters show significant differences by column at P ⬍ 0.05.

SFA, saturated fatty acids; MUSA, monounsaturated fatty acids; PUFA, polyunsaturated fatty acids;
w-3, total omega-3 fatty acids; w-6, total omega-6 fatty acids; EPA, eicosapentaenoic acid; DHA,
docosahexaenoic acid; SD, standard deviation of the mean.

tissue reported by Kovacs and Ackman (1978) and 0.24 g/100 g of edible-
portion value (U.S. Department of Agriculture, Agricultural Research Service
2004a). The PUFA content for WP (0.29 g/100 g of tissue) was lower than the
0.41 g/100 g of edible-portion value (U.S. Department of Agriculture, Agri-
cultural Research Service 2004b) in part because of their higher values for
both eicosapentaenoic acid (EPA) and DHA. The EPA content was highest in
ATF and lowest in GG. Even though the lipid content of ATF is much higher
than PC, the DHA content was similar (P ⬍ 0.05) for these two species, which
were both higher than WP or GG. It is well established that PL fractions have
higher concentrations of highly unsaturated FAs, such as DHA and EPA, when
compared to the TAG fractions (Gurr 1999).
In Alaska, the harvest of WP and PC accounted for 68% of the total
harvest in 2000 (Crapo and Bechtel 2003). The fat content and composition of
these two species were similar on a 100-g basis with both providing substantial
amounts of DHA (175–208 mg/100 g tissue). However, the EPA content of
both species was much lower than the DHA content (79–85 mg/100 g tissue).
Because of the low fat content in GG (0.21%), a 100-g portion of fillet would
provide only 50 mg of DHA and 28 mg of EPA. The ratio of EPA/DHA was of
interest as many marine finfish oils have ratios below 1.0 while others are
greater than 1.0. Values from this study were 0.55, 0.38, 0.48 and 1.66 for GG,
PC, WP and ATF, respectively.
FA Composition
Table 4 presents the percent of individual FAs in the total FAs analyzed
for each set of samples. Percent of myristic acid (C14:0) was significantly
 LIPID ANALYSIS OF FISH FILLETS 29

TABLE 4.
FATTY ACID METHYL ESTER ANALYSIS (PERCENT TOTAL FATTY ACIDS)

Giant grenadier Arrow-tooth flounder Pacific cod Walleye pollock

(SD) (SD) (SD) (SD)

C14:0 2.21A (0.63) 5.56B (0.32) 1.22C (0.30) 1.57AC (0.18)

C16:0 17.22A (1.37) 17.72A (1.01) 16.74A (0.41) 21.09B (1.38)
C16:1w7 3.10A (0.82) 6.59B (0.51) 1.64C (0.33) 2.11AC (0.43)
C16:3w4 0.63A (0.06) 0.26B (0.01) 0.66A (0.08) 0.00C (0.00)
C17:1w9 0.92A (0.15) 1.96B (0.17) 0.26C (0.44) 1.87B (0.13)
C18:0 4.55A (0.19) 2.36B (0.15) 5.44A (0.23) 4.89A (1.11)
C18:1w9 ci 11.10A (0.58) 17.77B (0.71) 8.83C (0.94) 11.53A (1.47)
C18:1w7 7.74A (2.19) 8.05A (0.59) 5.06B (0.56) 6.90AB (0.43)
C18:2w6 ci 0.90AB (0.17) 1.00A (0.07) 0.77AB (0.16) 0.70B (0.13)
C18:3w3 0.24A (0.23) 0.45A (0.04) 0.00B (0.00) 0.00B (0.00)
C18:4w3 0.53A (0.15) 1.20B (0.04) 0.18A (0.31) 0.19A (0.33)
C20:1w11 1.07A (0.10) 1.14A (0.32) 1.12A (0.31) 1.31A (0.41)
C20:1w9 1.57AB (0.37) 1.82B (0.22) 0.94C (0.18) 1.15AC (0.29)
C20:4w6 2.70A (0.28) 0.59B (0.06) 3.11A (0.43) 1.79C (0.16)
C21:0 3.29A (1.02) 0.09B (0.15) 0.00B (0.00) 0.00B (0.00)
C20:4w3 0.22A (0.24) 0.64B (0.06) 0.00A (0.00) 0.00A (0.00)
C20:5w3 13.19A (1.21) 12.52A (0.80) 13.40A (1.08) 14.17A (1.41)
C22:1w11 0.96A (0.04) 2.16B (0.56) 0.00C (0.00) 0.26C (0.44)
C22:1w9 1.02A (0.32) 0.39B (0.06) 0.00C (0.00) 0.00C (0.00)
C22:5w3 1.66A (0.42) 2.92B (0.43) 1.50A (0.05) 0.00C (0.00)
C22:6w3 23.69A (1.34) 7.53B (1.14) 35.18C (1.27) 29.22D (2.16)
S FAME ID 98.51A (0.67) 91.72A (6.07) 96.05A (2.52) 98.75A (2.09)

Different superscript letters show significant differences by column at P ⬍ 0.05.

S FAME ID, sum of fatty acids methyl esters identified; SD, standard deviation of the mean.

higher in ATF than other species. The percent palmitic acid (C16:0) was
highest in WP (P ⬎ 0.05) at over 20% of the total FAs in the extracted
lipids. This result was not surprising because C16:0 is a predominant FA in
fish lipids from a variety of waters around the world (Gruger et al. 1964).
The C16:0 content determined for PC fillets was slightly lower than the
18.2% determined by Stansby (1976). ATF had the highest percent of palmi-
toleic acid (C16:1w7) and PC the lowest. Percent stearic acid (C18:0) is
significantly lower in ATF than in other species, where values ranged from
4.5 to 5.4%. Values for C18:0 are in general agreement with those of
Stansby (1976), who reported PC fillet values of 4.7%. There were large
differences found in percent C18:1w9 among the species studied. ATF had
the most (17.8%) and PC the least (8.8%). C18:1w7 was significantly lower
in PC than in other fillets and both GG and ATF had higher percentages of
7.7 and 8.1%, respectively. All species had percent linoleic acid cis isomer
(C18:2w6) below 1%. However, ATF contained 0.5% a-linolenic acid
30 A.C.M. OLIVEIRA and P.J. BECHTEL

(C18:3w3), and GG about 0.2%. In the other two species, the signal for the
methyl ester of linolenic acid was below the limits for peak integration. ATF
had significantly more stearidonic acid (C18:4w3) at 1.2%, while the other
fillets had less than 0.5%. Gadoleic acid (C20:1w11) was not significantly
different among the species studied, while percent gondoic acid (C20:1w9)
was lowest in the extracted oils from PC and highest in ATF. Both PC and
GG had the most of arachidonic acid (AA) (C20:4w6), as a percent of the
total FAs in the fillets with values ranging from 3.1 to 2.7%, respectively.
Percent AA found in the other two species was below 2.0%. Ackman (1990)
reported that percent AA in most fish oils from northern latitude species was
below 0.5%. He also commented that in the human diet, AA has an antago-
nistic effect to the well known health benefits derived from a high w-3 FA
diet. However, the values determined herein were well within the range
reported for AA in a variety of marine fish species (Gruger et al. 1964).
Heneicosanoic acid (C21:0) was found in negligible amounts in the fillets of
all fish species except GG. The percent of this FA was surprisingly high,
averaging about 3.3%. Retention times for this peak precisely match those
for the C21:0 methyl ester. However, because this FA is not commonly found
in most fish species, further investigation using GC coupled with mass spec-
trometry is needed to confirm the identity of this peak. EPA or C20:5w3 was
not significantly different among species and was present in amounts over
12.5%. Percent cetoleic acid (C22:1w11) was significantly higher in ATF
(2.2%) than in all other species (less than 1%). Erucic acid (C22:1w9) was
not detected in PC or WP, and values in GG were 1%. Percent DPA or
C22:5w3 was highest in ATF (2.92%); however, the amount in WP was
below the limits of detection. There was a wide range of percent DHA or
C22:6w3 values among species. PC had the highest percent at about 35%,
which was higher than the 29% reported by Stansby (1976). ATF had a low
percent DHA, which was similar to the 7.9% value reported for Pacific
halibut (Gruger et al. 1964).
When the analysis for cod and pollock are compared, there are minor
differences in the FAME content, with the most notable being the absence of
C22:1w11 in cod and C22:5w3 in pollock fillets. Composition of the oil from
GG was similar to the FAME contents of both cod and pollock; however,
both C21:0 and C22:1w9 were found in much higher concentrations than in
cod, pollock or flounder. ATF had a fillet lipid content over five times greater
than those of the other three species (Table 1) and the most unique FA com-
position (Table 4). The percent of total FAs for C14:0, C16:1w7, C18:4w3
and C22:1w11 from ATF fillets were over twice as great as those from the
other species. However, the content of both C20:4w6 and C22:6w3 were over
three times lower than comparable values from fillets from any of the other
species.
 LIPID ANALYSIS OF FISH FILLETS 31

In this study, the lipids from PC, WP, GG and ATF fillets have been
extensively characterized. Results indicate that the nutritional value of the
lipids from PC and WP fillets were similar and would provide approximately
250–300 mg of w-3 FAs per 100 g of tissue. Although there is a huge
biomass of ATF in Alaskan waters; ATF is an underutilized species primarily
because of a very active protease that can result in poor texture properties of
the cooked flesh. In this study, it is shown that ATF has a relatively high
content of w-3 FAs, which would be of nutritional value and provide poten-
tial reasons to further utilize ATF fillets. From a processing perspective, ATF
can be automatically filleted and frozen or used to make ATF surimi. In
addition, the unique properties of ATF oil may have other food or industrial
applications. GG is also an underutilized species, although it had a much
lower w-3 FA content per 100 g of tissue. Further studies are needed to
identify products that could be made for this species to increase its
utilization.

CONCLUSIONS

GG is an underutilized species and its fillets had a low lipid content

(0.2%); however, ATF is also an underutilized species and its fillets contain a
much higher lipid content (3.5%). The ATF lipid had many unique properties,
including a low P/S ratio and high EPA/DHA ratio. WP and PC fillets had
similar lipid contents of approximately 0.6%. However, a number of differ-
ences were detected, including P/S, % total w-3 fatty acids/total % w-6 fatty
acids (w-3/w-6) and EPA/DHA ratios. There are several implications of the
findings of this study. The first is that there is a relatively high fat content in
ATF fillets (3.5%); consumption of 100 g of fillet would provide over 1 g of
MUSA and PUFA. In addition, the AFT w-3 FAs are somewhat unique in that
they have a low percent DHA, an average percent EPA and elevated percent-
ages of other w-3 FAs. These results would support increased consumption of
ATF and thus further its utilization. Another finding of this study is that GG
fillet contains a very low level of lipid (0.2%); a 100-g portion contributes less
than 100 mg of total w-3 FAs and thus would not be an obvious source of
dietary w-3 FAs.

ACKNOWLEDGMENTS

We thank the U.S. Department of Agriculture–Agricultural Research

Service and the University of Alaska Fairbanks Fishery Industrial Technology
Center for providing funds to this study. We also thank Jennifer Hoffert for her
technical expertise.
32 A.C.M. OLIVEIRA and P.J. BECHTEL

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